User:Jacob Esenther/Notebook/Chem 571/2014/09/10: Difference between revisions
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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span> | |style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==Objective== | ==Objective== | ||
# Explore the use of Bradford Analysis with Lysozyme | |||
==Description== | ==Description== | ||
# | # Bradford Analysis | ||
## Prepared 50 mL of a standard saline solution (0.9 wt-% NaCl). | |||
## Prepared 50 mL of a 50 mM Tris (not Tris-HCl) 50 mM NaCl solution. | |||
## Prepared a stock solution of '''BSA''' that is roughly 5 mg in 5 mL of saline. | |||
## Calculated your actual solution concentration. | |||
## Using a quartz cuvette, recorded UV-VIS spectra between 200 nm and 800 nm. | |||
## Prepared serial dilutions of 1μg/mL, 2μg/mL, 2.5μg/mL, 5μg/mL, 8μg/mL, 10μg/mL, 15μg/mL and 20μg/mL | |||
## Added 20μL of the serial dilutions to a 1.5mL centrifuge tube | |||
## Added 200μL of the Bio-Rad Protein Assay reagent. Use 1:4 concentrate diluted with water. | |||
## Added 780μL of Tris/NaCl buffer to make the final volume 1mL. | |||
## Obtained a UV-VIS spectrum. | |||
##* Blanks of Tris/NaCl buffer and 200 μLBradford reagent + 800μL buffer were also run | |||
==Data== | ==Data== | ||
[[Image:Bradford_Assay_of_Lysozyme.png|800x497px|center|thumb|Bradford Assay with Lysozyme]] | |||
[[Image:Bradford_Absorbance_v_Concentration.png|800x497px|center|thumb|Bradford Absorbance vs. Concentration]] | |||
[[Image:Bradford_Absorbance_v_Concentration_1_to_10.png|800x497px|center|thumb|Bradford Absorbance vs. Concentration (outliers removed)]] | |||
[[Image:Bradford_Stock_Solutions.png|800x497px|center|thumb|BSA and Lysozyme Stock Solutions for Bradford]] | |||
[[Image:Bradford_Corrected_BSA_Absorbance.png|800x497px|center|thumb|Corrected Bradford-BSA Absorbance]] | |||
==Notes== | ==Notes== | ||
No notes to report | |||
[[Category:Course]] | [[Category:Course]] |
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Objective
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DataNotesNo notes to report
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